Uncovering a role for micro-RNAs in sleep homeostasis and energy metabolism

Micro-RNAs (miRNAs) are key, post-transcriptional regulators of gene expression and have been implicated in almost every cellular process investigated thus far. However, their role in sleep, in particular the homeostatic aspect of sleep control, has received little attention. We here assessed the effects of sleep deprivation on the brain miRNA transcriptome in the mouse. Sleep deprivation affected miRNA expression in a brain-region specific manner. The forebrain expression of the miRNA miR-709 was affected the most and in situ analyses confirmed its robust increase throughout the brain, especially in the cerebral cortex and the hippocampus. The hippocampus was a major target of the sleep deprivation affecting 37 miRNAs compared to 52 in the whole forebrain. Moreover, independent from the sleep deprivation condition, miRNA expression was highly region-specific with 45% of all expressed miRNAs showing higher expression in hippocampus and 55% in cortex. Next we demonstrated that down-regulation of miRNAs in Com/c2o-expressing neurons of adult mice, through a conditional and inducible Dicer knockout mice model (cKO), results in an altered homeostatic response after sleep deprivation eight weeks following the tamoxifen-induced recombination. Dicer cKO mice showed a larger increase in the electro-encephalographic (EEG) marker of sleep pressure, EEG delta power, and a reduced Rapid Eye Movement sleep rebound, compared to controls, highlighting a functional role of miRNAs in sleep homeostasis. Beside a sleep phenotype, Dicer cKO mice developed an unexpected, severe obesity phenotype associated with hyperphagia and altered metabolism. Even more surprisingly, after reaching maximum body weight 5 weeks after tamoxifen injection, obese cKO mice spontaneously started losing weight as rapidly as it was gained. Brain transcriptome analyses in obese mice identified several obesity-related pathways (e.g. leptin, somatostatin, and nemo-like kinase signaling), as well as genes involved in feeding and appetite (e.g. Pmch, Neurotensin). A gene cluster with anti-correlated expression in the cerebral cortex of post-obese compared to obese mice was enriched for synaptic plasticity pathways. While other studies have identified a role for miRNAs in obesity, we here present a unique model that allows for the study of processes involved in reversing obesity. Moreover, our study identified the cortex as a brain area important for body weight homeostasis. Together, these observations strongly suggest a role for miRNAs in the maintenance of homeostatic processes in the mouse, and support the hypothesis of a tight relationship between sleep and metabolism at a molecular - Les micro-ARNS (miARNs) sont des régulateurs post-transcriptionnels de l'expression des gènes, impliqués dans la quasi-totalité des processus cellulaires. Cependant, leur rôle dans la régulation du sommeil, et en particulier dans le maintien de l'homéostasie du sommeil, n'a reçu que très peu d'attention jusqu'à présent. Dans cette étude, nous avons étudié les conséquences d'une privation de sommeil sur l'expression cérébrale des miARNs chez la souris, et observé des changements dans l'expression de nombreux miARNs. Dans le cerveau antérieur, miR-709 est le miARN le plus affecté par la perte de sommeil, en particulier dans le cortex cérébral et l'hippocampe. L'hippocampe est la région la plus touchée avec 37 miARNs changés comparés à 52 dans le cerveau entier. Par ailleurs, indépendamment de la privation de sommeil, certains miARNs sont spécifiquement enrichis dans certaines aires cérébrales, 45% des miARNs étant surexprimés dans l'hippocampe contre 55% dans le cortex. Dans une seconde étude, nous avons observé que la délétion de DICER, enzyme essentielle à la biosynthèse des miARNs, et la perte subséquente des miARNs dans les neurones exprimant la protéine CAMK2a altère la réponse homéostatique à une privation de sommeil, 8 semaines après l'induction de la recombinaison génétique par le tamoxifen. Les souris sans Dicer (cKO) ont une plus large augmentation de l'EEG delta power, le principal marqueur électro-encéphalographique du besoin de sommeil, comparée aux contrôles, ainsi qu'un rebond en sommeil paradoxal plus petit. De façon surprenante, les souris Dicer cKO développent une obésité rapide, sévère et transitoire, associée à de l'hyperphagie et une altération de leur métabolisme énergétique. Après avoir atteint un pic maximal d'obésité, les souris cKO entrent spontanément dans une période de perte de poids rapide. L'analyse du transcriptome cérébral des souris obèses nous a permis d'identifier des voies associées à l'obésité (leptine, somatostatine et nemo-like kinase), et à la prise alimentaire (Pmch, Neurotensin), tandis que celui des souris post-obèses a révélé un groupe de gènes liés à la plasticité synaptique. Au-delà des nombreux modèles d'obésité existant chez la souris, notre étude présente un modèle unique permettant d'étudier les mécanismes sous-jacent la perte de poids. De plus, nous avons mis en évidence un rôle important du cortex cérébral dans le maintien de la balance énergétique. En conclusion, toutes ces observations soutiennent l'idée que les miARNs sont des régulateurs cruciaux dans le maintien des processus homéostatiques et confortent l'hypothèse d'une étroite relation moléculaire entre le sommeil et le métabolisme.

Extent of Hypoattenuation On CT Angiography Source Images In Basilar Artery Occlusion: Prognostic Accuracy Of Posterior Circulation ASPECTS And The Pons-midbrain Score In The Basilar Artery International Cooperation Study

Background: The posterior circulation Acute Stroke Prognosis Early CT Score (pc-ASPECTS) and the combined Pons-midbrain score quantify the extent of early ischemic changes in the posterior circulation. We compared the prognostic accuracy of both scores if applied to CT angiography (CTA) source images (CTA-SI) of patients in the Basilar Artery International Cooperation Study (BASICS).Methods: BASICS was a prospective, observational, multi-centre, registry of consecutive patients who presented with acute symptomatic basilar artery occlusion (BAO). Functional outcome was assessed at 1 month. We applied pc-ASPECTS and the combined Pons-midbrain score to CTA-SI by 3-reader-consensus. Readers were blinded to clinical data. We performed multivariable logistic regression analysis, adjusting for thrombolysis, baseline NIHSS score and age, and used the output to derive ROC curves to compare the ability of both scores to discriminate patients with favourable (modified Rankin Scale [mRS] scores 0-3) from patients with unfavourable (mRS scores 4-6) functional outcome.Results: We reviewed CTAs of 158 patients (64% men, mean age 65 _ 15 years, median NIHSS score 25 [0-38], median GCS score 7 [3-15], median onset-to-CTA time 234 minutes [11-7380]). At 1 month, 40 (25%) patients had a favourable outcome, 49 (31%) had an unfavourable outcome (mRS score 4-5) and 69 (44%) were deceased. Both techniques of assessing CTA-SI hypoattenuation in the posterior circulation showed equally good discriminative value in predicting final outcome (C-statistics; area under ROC curve 0.74 versus 0.75, respectively; p_0.37). Pc-ASPECTS dichotomized at _6 versus _6 was an independent predictor of favourable functional outcome (RR _ 2.2; CI95 1.1-4.7; p _ 0.034).Conclusion: Compared to the combined Pons-midbrain score, the pc-ASPECTS score has similar prognostic accuracy to identify patients with a favourable functional outcome in BASICS. Dichotomized pc-ASPECTS (_6 versus _6) is an independent predictor of favourable functional outcome in this population. Author Disclosures: V. Puetz: None. A. Khomenko: None. M.D. Hill: None. I. Dzialowski: None. P. Michel: None. C. Weimar: None. C.A.C. Wijman: None. H. Mattle: None. K. Muir: None. T. Pfefferkorn: None. D. Tanne: None. S. Engelter: None. K. Szabo: None. A. Algra: None. A.M. Demchuk: None. W.J. Schonewille: None.

Volumetric quantification of atherosclerotic plaque in CT considering partial volume effect.

Coronary artery calcification (CAC) is quantified based on a computed tomography (CT) scan image. A calcified region is identified. Modified expectation maximization (MEM) of a statistical model for the calcified and background material is used to estimate the partial calcium content of the voxels. The algorithm limits the region over which MEM is performed. By using MEM, the statistical properties of the model are iteratively updated based on the calculated resultant calcium distribution from the previous iteration. The estimated statistical properties are used to generate a map of the partial calcium content in the calcified region. The volume of calcium in the calcified region is determined based on the map. The experimental results on a cardiac phantom, scanned 90 times using 15 different protocols, demonstrate that the proposed method is less sensitive to partial volume effect and noise, with average error of 9.5% (standard deviation (SD) of 5-7mm(3)) compared with 67% (SD of 3-20mm(3)) for conventional techniques. The high reproducibility of the proposed method for 35 patients, scanned twice using the same protocol at a minimum interval of 10 min, shows that the method provides 2-3 times lower interscan variation than conventional techniques.

Solitary fibrous tumor of the lateral ventricle: CT appearances and pathologic correlation with follow-up.

Intracranial solitary fibrous tumors are rare, and intraventricular fibrous tumors are even more unusual. We report a case of solitary fibrous tumor in the region of trigone and body of the left lateral ventricle and discuss the clinical presentation, CT characteristics, and histopathologic features with 1-year follow-up. We speculate that the tumor arose from the perivascular connective tissue of the choroid plexus.

Benign Intrapulmonary Schwannoma: Aspect on F-18 Fluorodeoxyglucose PET/CT.

A 75-year-old man, with no significant symptoms, was referred after the incidental finding of a left hilar pulmonary mass of 30 × 30 × 50 mm on a chest CT. F-18 fluorodeoxyglucose (FDG) PET/CT demonstrated a heterogeneous, moderate radiotracer uptake in the mass (SUV 3.5 g/mL). Bronchoscopy revealed a discrete extrinsic compression of the superior bronchus without endobronchial lesion. Endobronchial fine-needle biopsies could not deliver a final diagnosis. The patient underwent upper lobectomy by thoracotomy. Histopathology revealed a benign intrapulmonary schwannoma. Although rare, intermediate FDG uptake in the settings of a pulmonary mass should include schwannoma in the differential diagnosis.

Etude pilote par PET/CT de la réponse à l'ipilimumab dans le traitement du mélanome métastatique.

0 Abstract L'incidence du mélanome est en nette augmentation en Europe et aux Etats-Unis. La Chirurgie peut être curative au stade précoce de la maladie,la radiothérapie se pratique À visée palliative, les chimiothérapies n'ont montré que peu d'effet. Les progress actuels se font via l'immunothérapie. Des traitements par l'interleukine-2 et l'interféron-α se sont montrés efficaces chez certains patients, mais leur utilisation est limitée par leur forte toxicité. Depuis 2011, en Suisse, une nouvelle molécule a été mise sur le marché, l'ipilimumab (Yervoy®). Il s'agit d'un anticorps! monoclonal humain dirigé contre le CTLA4. Il en résulte une activation non spécifique du système immunitaire. Une étude randomisée de phase IIIa été conduite au Etats-Unis. Elle démontre une augmentation de la survie chez les patients atteints de melanoma métastatique traits par ipilimumab, sans traitement préalable des métastases. Le but de cette etude est de determiner s'il est possible, par le biais de l'imagerie PET-CTau 18F-FDG, de prédire la réponse individuelle au traitement par ipilimumab, afin d'optimiser la prise en charge de ces patients. 0.1 Méthode Les patients atteints de mélanome métastatique sont exposés au traitement d'ipilimumab selon les recommandations de Bristol-Myers Squibb. Puis ils effectuent des PET-CT au F-18-FDG selon! le! protocole (CER 400/11, annexé) de l'étude. Les images sont!analysées selon les critères PERCIST (PETResponse Criterias In Solid Tumors) et le TLG (Total Lesion Glycolysis) est calculé. Parallèlement, des prises de sang sont effectuées et les échantillons sont analysés à l'institut Ludwig (LICR, Unil) selon les critères imRC qui determinant la réponse immunologique au traitement. 0.2 Résultats Nous notons une discrépance dans les résultats. Lorsque nous observons une maladie stable avec les critères immunologiques imRC, nous observons une maladie progressive avec lescritères PERCIST. 0.3 Discussion - Conclusion Nous n'avons pu faire entrer que cinq patients dans l'étude, dont trois étaient vivants à trois mois, ce qui a restreint le nombre de données analysables. Les discrépances que nous observons dans nos résultats pourraient être dues au fait que le PET-CT au 18FDG ne nous permet pas de différencier l'activité tumorale de l'activité inflammatoire péri-tumorale. Ce biais pourrait être à l'avenir prévenu en utilisant du 18FLT, un marqueur plus sélectif des cellules tumorales.L'analyse desimages avec les critères de total-lesion glycolysis a été impossible au vu du nombre de lésions que présentaient certains patients. Les critères PERCIST exigent un protocole d'acquisition très strict. La corrélation entre les images, les données immunologiques et la clinique mérite d'être suivie sur le long terme, car il pourrait y avoir une réponse positive plus tardive du traitement par ipilimumab.

Relation between intraoperative EMG values and final pedicle screw position as seen on CT images

Introduction: Intraoperative EMG based neurophysiological monitoring is increasingly used to assist pedicle screw insertion. We carried out a study comparing the final screw position in the pedicle measured on CT images in relation to its corresponding intraoperative muscle compound action potential (CMAP) values. Material and methods: A total of 189 screws were inserted in thoracolumbar spines of 31 patients during instrumented fusion under EMG control. An observer, blinded to the CMAP value, assessed the horizontal and vertical 'screw edge to pedicle edge' distance perpendicular to the longitudinal axis of the screw on reformatted CT reconstructions using OsiriX software. These distances were analysed with their corresponding CMAP values. Data from 62 thoracic and 127 lumbar screws were processed separately. Interobserver reliability of distance measurements was assessed. Results: No patient suffered neurological injury secondary to screw insertion. Distance measurements were reliable (paired t-test, P = 0.13/0.98 horizontal/vertical). Two screws had their position altered due to low CMAP values suggesting close proximity of nerve tissue. Seventy five percent of screws had CMAP results above 10mA and had an average distance of 0.35cm (SD 0.23) horizontally and 0.46cm (SD 0.26) vertically from the pedicle edge. Additional 12% had a distance from the edge of the pedicle less than 0mm indicating cortical breach but had CMAP values above 10mA. A poor correlation between CMAP values and screw position was found. Discussion: In this study CMAP values above 10mA indicated correct screw position in the majority of cases. The zone of 10-20mA CMAP carries highest risk of a misplaced screw despite high CMAP value (17% of screws this CMAP range). In order to improve accuracy of EMG predictive value further research is warranted including improvement of probing techniques.

Bone mineral density (BMD), micro-architecture estimation (TBS) and vertebral fracture assessment (VFA) extracted from a single DXA device in combination with clinical risk factors improve significantly the identification of women at high risk of fracture

Introduction: Osteoporosis (OP) is a systemic skeletal disease characterized by a low bone mineral density (BMD) and a micro-architectural (MA) deterioration. Clinical risk factors (CRF) are often used as a MA approximation. MA is yet evaluable in daily practice by the Trabecular Bone Score (TBS) measure. TBS is a novel grey-level texture measurement reflecting bone micro-architecture based on the use of experimental variograms of 2D projection images. TBS is very simple to obtain, by reanalyzing a lumbar DXA-scan. TBS has proven to have diagnosis and prognosis value, partially independent of CRF and BMD. The aim of the OsteoLaus cohort is to combine in daily practice the CRF and the information given by DXA (BMD, TBS and vertebral fracture assessment (VFA)) to better identify women at high fracture risk. Method: The OsteoLaus cohort (1400 women 50 to 80 years living in Lausanne, Switzerland) started in 2010. This study is derived from the cohort COLAUS who started in Lausanne in 2003. The main goals of COLAUS is to obtain information on the epidemiology and genetic determinants of cardiovascular risk in 6700 men and women. CRF for OP, bone ultrasound of the heel, lumbar spine and hip BMD, VFA by DXA and MA evaluation by TBS are recorded in OsteoLaus. Preliminary results are reported. Results: We included 631 women: mean age 67.4±6.7 y, BMI 26.1±4.6, mean lumbar spine BMD 0.943±0.168 (T-score -1.4 SD), TBS 1.271±0.103. As expected, correlation between BMD and site matched TBS is low (r2=0.16). Prevalence of VFx grade 2/3, major OP Fx and all OP Fx is 8.4%, 17.0% and 26.0% respectively. Age- and BMI-adjusted ORs (per SD decrease) are 1.8 (1.2- 2.5), 1.6 (1.2-2.1), 1.3 (1.1-1.6) for BMD for the different categories of fractures and 2.0 (1.4-3.0), 1.9 (1.4-2.5), 1.4 (1.1-1.7) for TBS respectively. Only 32 to 37% of women with OP Fx have a BMD < -2.5 SD or a TBS < 1.200. If we combine a BMD < -2.5 SD or a TBS < 1.200, 54 to 60% of women with an osteoporotic Fx are identified. Conclusion: As in the already published studies, these preliminary results confirm the partial independence between BMD and TBS. More importantly, a combination of TBS subsequent to BMD increases significantly the identification of women with prevalent OP Fx which would have been miss-classified by BMD alone. For the first time we are able to have complementary information about fracture (VFA), density (BMD), micro- and macro architecture (TBS & HAS) from a simple, low ionizing radiation and cheap device: DXA. Such complementary information is very useful for the patient in the daily practice and moreover will likely have an impact on cost effectiveness analysis.